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You searched for: EV210035 (EV-TRACK ID)
Showing 1 - 2 of 2
Showing 1 - 2 of 2
Details | EV-TRACK ID | Experiment nr. | Species | Sample type | Separation protocol | First author | Year | EV-METRIC |
---|---|---|---|---|---|---|---|---|
EV210035 | 2/2 | Homo sapiens | A-431 |
(d)(U)C DG Filtration |
van Dommelen, Susan M | 2016 | 78% | |
Study summaryFull title
All authors
Susan M van Dommelen, Roy van der Meel, Wouter W van Solinge, Maria Coimbra, Pieter Vader, Raymond M Schiffelers
Journal
Nanomedicine
Abstract
Aim: Extracellular vesicles (EVs) are attractive candidates for biomarker research, because their co (show more...)
EV-METRIC
78% (97th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Cetuximab
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Density gradient Filtration Protein markers
EV: TSG101/ Alix/ CD9/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ Cetuximab
non-EV: Lamin-A/ Lamin-C/ ATP5-A/ Tom20 Proteomics
no
EV density (g/ml)
1.10-1.21
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
A-431
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: rotor type
JA-30.50
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
Not specified
Wash: time (min)
70
Wash: Rotor Type
JA-30.50
Wash: speed (g)
100000
Density gradient
Only used for validation of main results
Yes
Type
Continuous
Lowest density fraction
0.4
Highest density fraction
2.5
Total gradient volume, incl. sample (mL)
Not specified
Sample volume (mL)
Not spec
Orientation
Top-down
Rotor type
SW 40 Ti
Speed (g)
200000
Duration (min)
960
Fraction volume (mL)
1
Fraction processing
Centrifugation
Pelleting: volume per fraction
4
Pelleting: duration (min)
70
Pelleting: rotor type
Not specified
Pelleting: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
CD9/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ Cetuximab/ TSG101/ Alix
Not detected contaminants
Lamin-A/ Lamin-C/ ATP5-A/ Tom20
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Not Reported
EV concentration
Yes
|
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EV210035 | 1/2 | Homo sapiens | A-431 |
(d)(U)C Filtration |
van Dommelen, Susan M | 2016 | 56% | |
Study summaryFull title
All authors
Susan M van Dommelen, Roy van der Meel, Wouter W van Solinge, Maria Coimbra, Pieter Vader, Raymond M Schiffelers
Journal
Nanomedicine
Abstract
Aim: Extracellular vesicles (EVs) are attractive candidates for biomarker research, because their co (show more...)
EV-METRIC
56% (90th percentile of all experiments on the same sample type)
Reported
Not reported Not applicable EV-enriched
proteins
Protein analysis: analysis of three or more EV-enriched proteins
non
EV-enriched protein
Protein analysis: assessment of a non-EV-enriched protein
qualitative
and quantitative analysis
Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected.
electron
microscopy images
Particle analysis: inclusion of a widefield and close-up electron microscopy image
density
gradient
Separation method: density gradient, at least as validation of results attributed to EVs
EV density
Separation method: reporting of obtained EV density
ultracentrifugation specifics
Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps
antibody
specifics
Protein analysis: antibody clone/reference number and dilution
lysate
preparation
Protein analysis: lysis buffer composition
Study dataSample type
Cell culture supernatant
Sample origin
Control condition
Focus vesicles
extracellular vesicle
Separation protocol
Separation protocol
(Differential) (ultra)centrifugation
Filtration Protein markers
EV: TSG101/ Alix/ Cetuximab/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ CD9
non-EV: Lamin-A/ Lamin-C/ ATP5-A/ Tom20 Proteomics
no
Show all info
Study aim
Function/Identification of content (omics approaches)
Sample
Species
Homo sapiens
Sample Type
Cell culture supernatant
EV-producing cells
A-431
EV-harvesting Medium
Serum free medium
Separation Method
(Differential) (ultra)centrifugation
dUC: centrifugation steps
Below or equal to 800 g
Between 800 g and 10,000 g Between 100,000 g and 150,000 g Pelleting performed
Yes
Pelleting: time(min)
70
Pelleting: rotor type
JA-30.50
Pelleting: speed (g)
100000
Wash: volume per pellet (ml)
Not specified
Wash: time (min)
70
Wash: Rotor Type
JA-30.50
Wash: speed (g)
100000
Filtration steps
0.22µm or 0.2µm
Characterization: Protein analysis
Protein Concentration Method
microBCA
Western Blot
Detected EV-associated proteins
CD9/ EGFR/ pEGFR/ Akt/ pAkt/ -actin/ TSG101/ Alix
Not detected EV-associated proteins
Cetuximab
Not detected contaminants
Lamin-A/ Lamin-C/ ATP5-A/ Tom20
Characterization: Lipid analysis
No
Characterization: Particle analysis
NTA
Report type
Not Reported
EV concentration
Yes
|
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1 - 2 of 2 |
EV-TRACK ID | EV210035 | |
---|---|---|
species | Homo sapiens | |
sample type | Cell culture | |
cell type | A-431 | |
condition | Cetuximab | Control condition |
separation protocol | dUC Density gradient Filtration | dUC Filtration |
Exp. nr. | 2 | 1 |
EV-METRIC % | 78 | 56 |